Docosapentaenoic acids (DPAs) are long chain polyunsaturated fatty acids that exist as two major structural isomers: n-3 DPA and n-6 DPA. n-3 DPA is found in seal meat, salmon and abalone, and n-6 DPA is found in several marine microbial oil. We investigated the bioconversion of n-3 and n-6 DPAs in three different human cell lines, Caco-2, HepG2, and THP-1. n-3 DPA was converted to docosahexaenoic acid only in HepG2 cells. In contrast, retro-conversion to eicosapentaenoic acid (EPA) was observed in all three cell lines. n-6 DPA was also retro-converted to arachidonic acid (AA) in Caco-2 and HepG2 cells. EPA and AA were particularly elevated in Caco-2 cells, compared to HepG2 cells. Further, the retro-conversion of n-3 DPA led to a greater increase of EPA in the phospholipid fraction than in the neutral lipid fraction.